Sustained-release dosage forms of ruxolitinib

ABSTRACT

The present invention relates to sustained-release formulations and dosage forms of ruxolitinib, or a pharmaceutically acceptable salt thereof, which are useful in the treatment of Janus kinase-associated diseases such as myeloproliferative disorders.

FIELD OF THE INVENTION

The present invention relates to sustained-release formulations anddosage forms of ruxolitinib, or a pharmaceutically acceptable saltthereof, which are useful in the treatment of Janus kinase-associateddiseases such as myeloproliferative disorders.

BACKGROUND OF THE INVENTION

Ruxolitinib((3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propanenitrile)is the first FDA approved Janus kinase (JAK) inhibitor and is the onlydrug currently approved for treatment of myelofibrosis. Mascarenhas, J.et al. Clin Cancer Res. 2012 Jun. 1; 18(11):3008-14. Epub 2012 Apr. 2.The compound has been shown in the clinic to effectively reduce spleenvolume and improve total symptom scores in patients suffering frommyelofibrosis. See, e.g., Verstovsek, S., et al. “A double-blind,placebo-controlled trial of ruxolitinib for myelofibrosis,” N. Eng. J.Med., 2012, Mar. 1:366(9):799-807, which is incorporated herein byreference in its entirety, which reports the results of a Phase 3clinical trial (COMFORT-I Study) of ruxolitinib for myelofibrosis. Seealso, Harrison, C. et al., “JAK inhibition with ruxolitinib versus bestavailable therapy for myelofibrosis,” N. Eng. J. Med., 2012, Mar. 1;366(9):787-98 reporting Phase 3 clinical trial results of the COMFORT-IIstudy, which is incorporated herein by reference in its entirety.

To date, all published human clinical data for ruxolitinib relate todosing of an immediate-release formulation. However, ruxolitinib is aBCS Class I molecule with rapid oral absorption and a short half-life ofabout 3 hours. See, Shi et al., J. Clin. Pharmacol. 2012 June;52(6):809-18. Epub 2011 May 20. These properties result in a highpeak/trough plasma concentration ratio in human subjects leading tomultiple daily doses for optimal treatment, and potentially contributingto problems with patient compliance and unwanted side effects.

Ruxolitinib therapy is often associated with the adverse events ofthrombocytopenia (low platelet count) and anemia (low hemoglobin).Thrombocytopenia is dose-dependent and considered the dose-limitingtoxic effect.

Accordingly, there is a need for new and improved formulations ofruxolitinib that not only mitigate adverse side-effects in patients, butstill achieve therapeutic efficacy, and also facilitate administrationof the drug such as by reducing the number of doses required to achievea therapeutic effect. The sustained-release formulations provided hereinhelp meet these and other needs.

SUMMARY OF THE INVENTION

The present invention is directed to a sustained-release dosage formcomprising at least one active ingredient which is ruxolitinib, or apharmaceutically acceptable salt thereof, wherein the ruxolitinib, orpharmaceutically acceptable salt thereof, is present in the dosage formin an amount of about 10 to about 60 mg on a free base basis.

The present invention is further directed to a method of treating adisease associated with JAK activity in a patient in need thereof,comprising administering the sustained-release dosage form of theinvention to said patient.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a graph comparing plasma concentrations of ruxolitinibafter administration of a single dose of either 25 mg immediate-releaseor 25 mg sustained-release formulation in fasted, healthy humansubjects.

FIG. 2 shows a graph comparing spleen volume responders in the COMFORT-Iimmediate-release formulation study and the sustained-release study.

FIG. 3 shows a graph comparing total symptom scores in the COMFORT-Iimmediate-release formulation study and the sustained-release study.

FIG. 4 shows a graph comparing ruxolitinib steady state plasmaconcentrations in MF patients dosed with 25 mg immediate-release or 25mg sustained-release formulations.

DETAILED DESCRIPTION Sustained-Release Dosage Forms

The present invention provides, inter alia, an oral, sustained-releasedosage form comprising ruxolitinib, or a pharmaceutically acceptablesalt thereof, as an active ingredient. The dosage form can containruxolitinib, or a pharmaceutically acceptable salt thereof, in an amountof about 10 to about 60 mg, about 10 to about 40 mg, about 20 to about40 mg, or about 20 to about 30 mg on a free base basis. In someembodiments, the dosage form contains about 10 mg, about 12.5 mg, about20 mg, about 25 mg, about 30 mg, about 37.5 mg, about 40 mg, about 50mg, or about 60 mg on a free base basis. In some embodiments, the dosageform contains about 25 mg of ruxolitinib on a free base basis. Thephrase “on a free base basis” indicates that the amount of ruxolitinibor salt thereof in the dosage form is measured based on the molecularweight of ruxolitinib free base only, even when the actual activeingredient is a salt of ruxolitinib having a different molecular weightthan the free base. For example, the conversion factor for ruxolitinibphosphate salt to free base is 0.7575.

The structure, preparation, and characterization of ruxolitinib, andpharmaceutically acceptable salts thereof, are described in, e.g., U.S.Pat. No. 7,598,257 and US Pat. Pub. No. 2008/0312259, each of which isincorporated herein by reference in its entirety. In some embodiments,the active ingredient is a pharmaceutically acceptable salt ofruxolitinib, such as the maleic acid salt, sulfuric acid salt, orphosphoric acid salt. In some embodiments, the active ingredient isruxolitinib phosphate (i.e., phosphoric acid salt of ruxolitinib).

The dosage form of the invention comprises a sustained-releaseformulation of ruxolitinib, or a pharmaceutically acceptable saltthereof. As used herein, “sustained-release” is used as generallyunderstood in the art and refers to a formulation designed to slowlyrelease the active ingredient into a patient after oral administrationand to maintain an essentially steady, therapeutically effective plasmalevel of active ingredient over a relatively long period of time, suchas about 8 to about 24 hours or longer.

The dosage forms of the invention include a sustained-release matrixformer. Example sustained-release matrix formers include cellulosicethers such as hydroxypropyl methylcellulose (HPMC, hypromellose) whichis a high viscosity polymer. The sustained-release dosage forms of theinvention can include, for example, about 10 to about 30%, about 15 toabout 25%, or about 18 to about 24% by weight of hydroxypropylmethylcellulose(s). In some embodiments, the formulation has about 20%by weight of one or more hydroxypropyl methylcelluloses. In furtherembodiments, the formulation has about 22% by weight of one or morehydroxypropyl methyl celluloses. Example hydroxypropyl methylcellulosesinclude Methocel K15M, Methocel K4M, and Methocel K100LV.

The sustained-release dosage forms of the invention can further includeone or more fillers, glidants, disintegrants, binders, or lubricants asinactive ingredients. Fillers can be present in the formulations in anamount of 0 to about 85% by weight. In some embodiments, the formulationhas about 50 to about 80%, about 55 to about 75%, or about 60 to about70% by weight of filler. Non-limiting examples of fillers includelactose monohydrate, microcrystalline cellulose, starch 1500, andlactose anhydrous, or combinations thereof. In some embodiments, thefiller comprises microcrystalline cellulose, lactose monohydrate, orboth.

Lubricants can be present in the dosage forms of the invention in anamount of 0 to about 5% by weight. Non-limiting examples of lubricantsinclude magnesium stearate, stearic acid (stearin), hydrogenated oil,polyethylene glycol, sodium stearyl fumarate, and glyceryl behenate. Insome embodiments, the formulations include magnesium stearate, stearicacid, or both.

Glidants can be present in the dosage forms of the invention in anamount of 0 to about 5% by weight. Non-limiting examples of glidantsinclude talc, colloidal silicon dioxide, and cornstarch. In someembodiments, the glidant is colloidal silicon dioxide.

Disintegrants can be present in the dosage forms of the invention in anamount of 0 to about 10% by weight. Non-limiting examples ofdisintegrants include croscarmellose sodium, crospovidone, starch,cellulose, and low substituted hydroxypropyl cellulose. Croscarmellosesodium is a preferred disintegrant.

Film-coating agents can be present in an amount of 0 to about 5% byweight. Non-limiting illustrative examples of film-coating agentsinclude hypromellose or polyvinyl alcohol based coating with titaniumdioxide, talc and optionally colorants available in several commerciallyavailable complete coating systems.

In some embodiments, the dosage form of the invention includes asustained-release formulation comprising about 12.2% ruxolitinibphosphate, about 20% hydroxypropyl methylcellulose, about 64.3% filler,about 2.5% lubricant, and about 1% glidant, all by weight.

In some embodiments, the dosage form of the invention includes asustained-release formulation comprising about 12.2% ruxolitinibphosphate, about 22% hydroxypropyl methylcellulose, about 62.3% filler,about 2.5% lubricant, and about 1% glidant, all by weight.

In some embodiments, the dosage form of the invention includes asustained-release formulation as set out below.

Percentage Component (wt %) Ruxolitinib phosphate 12.2 Microcrystallinecellulose, NF 22.0 Hypromellose, USP (Methocel K15M) 4.0 Hypromellose,USP (Methocel K4M) 16.0 Lactose monohydrate, NF 42.3 Colloidal silicondioxide, NF 1.0 Magnesium stearate, NF 0.5 Stearic acid, NF 2.0

In some embodiments, the dosage form of the invention includes asustained-release formulation as set out below.

Percentage Component (wt %) Ruxolitinib phosphate 12.2 Microcrystallinecellulose, NF 42.3 Hypromellose, USP (Methocel K100LV) 10.0Hypromellose, USP (Methocel K4M) 12.0 Lactose monohydrate, NF 20.0Colloidal silicon dioxide, NF 1.0 Magnesium stearate, NF 0.5 Stearicacid, NF 2.0

In some embodiments, the dosage form of the invention includes asustained-release formulation as set out below.

Percentage Component (wt %) Ruxolitinib phosphate 12.2  Microcrystallinecellulose, NF 22.0-42.3 Hypromellose, USP (Methocel K100LV)   0-10.0Hypromellose, USP (Methocel K15M)   0-4.0 Hypromellose, USP (MethocelK4M) 12.0-16.0 Lactose monohydrate, NF 20.0-42.3 Colloidal silicondioxide, NF 1.0 Magnesium stearate, NF 0.5 Stearic acid, NF 2.0

As used herein, the term “dosage form” is meant to refer to a physicallydiscrete unit of sustained-release formulation of the invention to beadministered to a patient. Example dosage forms include tablets,caplets, capsules, and the like, containing any of the sustained-releaseformulations described herein. Dosage forms can further includepharmaceutically acceptable coatings, pigments, or dyes.

The dosage forms of the invention contain a sustained-releaseformulation that results in the relatively slow release of ruxolitinibonce administered, characterized by particular pharmacokineticparameters different from those of an immediate-release formulation. Thesustained-release dosage forms of the invention can minimize potentiallyharmful spikes in drug plasma concentrations that are associated withimmediate-release formulations, and can help provide continuous, steady,and therapeutically effective plasma levels of drug. The dosage forms ofthe invention can be administered to a human patient as needed fortherapeutic efficacy against the disease being treated, for example,once daily.

In some embodiments, the dosage forms of the invention are administeredto fasted patients. As used herein, “fasted” means, in reference to ahuman patient or subject, that the patient or subject has not ingestedfood or drink (except water) for at least 3 hours prior to dosing. Insome embodiments, patients are fasted for at least 10 hours prior todosing.

In further embodiments, the dosage forms of the invention areadministered to non-fasted human patients or subjects. Bioavailabilityof ruxolitinib is high (e.g., about 70-80%) and no food effect has beenobserved in immediate-release dosage forms. Accordingly, it is believedthat the pharmacokinetics of ruxolitinib administered as asustained-release dosage form would not be significantly different infasted and non-fasted patients.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 700 nM or less.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 600 nM or less.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 500 nM or less.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 400 nM or less.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 200 to about 700 nM.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 200 to about 600 nM.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 300 to about 500 nM.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 300 to about 400 nM.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results a mean time to peak plasmaconcentration (T_(max)) of ruxolitinib of about 1.5 hours or more.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean time to peak plasmaconcentration (T_(max)) of ruxolitinib of about 1.5 hours to about 5hours.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean time to peak plasmaconcentration (T_(max)) of ruxolitinib of about 2 hours to about 4hours.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a ratio of mean peak plasmaconcentration (C_(max)) to mean 12-hour plasma concentration (C_(12h))of ruxolitinib of about 10 or less.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a ratio of mean peak plasmaconcentration (C_(max)) to mean 12-hour plasma concentration (C_(12h))of ruxolitinib of about 6 or less.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a ratio of mean peak plasmaconcentration (C_(max)) to mean 12-hour plasma concentration (C_(12h))of ruxolitinib of about 5 or less.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a ratio of mean peak plasmaconcentration (C_(max)) to mean 12-hour plasma concentration (C_(12h))of ruxolitinib of about 4 or less.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a ratio of mean peak plasmaconcentration (C_(max)) to mean 12-hour plasma concentration (C_(12h))of ruxolitinib of about 1 to 10.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a ratio of mean peak plasmaconcentration (C_(max)) to mean 12-hour plasma concentration (C_(12h))of ruxolitinib of about 2 to 7.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean half-life (t_(1/2)) offrom about 3.5 hours to about 11 hours.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a mean half-life (t_(1/2)) offrom about 4 hours to about 8 hours.

In some embodiments, administration of a single dose of asustained-release dosage form of the invention to a human results inmean bioavailability (AUC_(0-∞)) of ruxolitinib of at least about 3000nM*h.

In some embodiments, administration of a single dose of asustained-release dosage form of the invention to a human results inmean bioavailability (AUC_(0-∞)) of ruxolitinib of at least about 3500nM*h.

In some embodiments, administration of a single dose of asustained-release dosage form of the invention to a human results inmean bioavailability (AUC_(0-∞)) of ruxolitinib of about 3000 to about4000 nM*h.

In some embodiments, administration of a single dose of asustained-release dosage form of the invention to a human results inmean bioavailability (AUC_(0-∞)) of ruxolitinib of about 3100 to about3800 nM*h.

In some embodiments, the sustained-release dosage form of the inventionhas a mean relative bioavailability based on AUC of from about 65% toabout 110% or about 75% to about 95% relative to an immediate releaseformulation comprising the same amount of ruxolitinib, or apharmaceutically acceptable salt thereof, in patients. AUC can be, forexample, AUC_(0-∞) (e.g., for a single dose) or AUC_(0-t) where t is aspecified time.

As used herein, “mean” when preceding a pharmacokinetic value (e.g. meanC_(max)) represents the arithmetic mean value of the pharmacokineticvalue taken from a population of patients unless otherwise specified.

As used herein, “C.” means the maximum observed plasma concentration.

As used herein, “C_(12h)” refers to the plasma concentration measured at12 hours from administration.

As used herein, “T_(max)” refers to the time at which the maximum bloodplasma concentration is observed.

As used herein, “T_(1/2)” refers to the time at which the plasmaconcentration is half of the observed maximum.

As used herein, “AUC” refers to the area under the plasmaconcentration-time curve which is a measure of total bioavailability.

As used herein, “AUC_(0-∞)” refers to the area under the plasmaconcentration-time curve extrapolated to infinity.

As used herein, “AUC_(0-t)” refers to the area under the plasmaconcentration-time curve from time 0 to the last time point with aquantifiable plasma concentration, usually about 12-36 hours.

As used herein, “AUC_(0-t)” refers to the area under the plasmaconcentration-time curve from time 0 to the time of the next dose.

As used herein, “Cl/F” refers to oral clearance.

The sustained-release dosage forms of the invention have certainadvantages over immediate-release dosage forms. The maintenance ofsteady, therapeutically effective plasma levels of ruxolitinib affordedby the sustained-release dosage forms of the invention allows forreduced dosing, such as doing only once per day, as opposed to twice ormore for immediate-release forms. The reduced dosing can help withpatient compliance in their treatment regimen.

In some embodiments, administration of the sustained-release dosage formof the invention to a human results in a therapeutically effectiveplasma level of ruxolitinib for at least about 8 hours, at least about10 hours, at least about 12 hours, at least about 18, or at least about24 hours. In some embodiments, the sustained-release dosage form of theinvention maintains a plasma level between about 75 and about 500 nM forat least about 8 hours, at least about 12 hours, or at least about 18hours. In some embodiments, the sustained-release dosage form of theinvention maintains a plasma level between about 100 and about 400 nMfor at least about 6 hours or at least about 8 hours.

An additional advantage of the sustained-release dosage form (e.g.,containing 25 mg of ruxolitinib phosphate on a free base basis) includesa reduction in unwanted side effects related to thrombocytopenia andanemia while maintaining therapeutic efficacy comparable with animmediate-release dosing regimen, e.g., 15 mg or 20 mg BID. It was notpredictable that a sustained-release formulation of ruxolitinib couldboth maintain therapeutic efficacy and significantly reduce unwantedside effects related to thrombocytopenia or reduced hemoglobin levels.Clinical data related to efficacy and side effects in myelofibrosispatients for both sustained-release and immediate-release dosing iscompared in the Examples.

In some embodiments, administration of the sustained-release dosage formof the invention to a human once daily for at least 16 weeks results ina mean decrease in mean base platelet count of no more than about100×10⁹/L, no more than about 80×10⁹/L, no more than about 60×10⁹/L, orno more than about 40×10⁹/L.

In some embodiments, administration of the sustained-release dosage formof the invention to a human once daily for at least 16 weeks results ina mean decrease in mean base platelet count of between about 0×10⁹/L andabout 100×10⁹/L, between about 30×10⁹/L and about 80×10⁹/L, or betweenabout 50×10⁹/L and about 70×10⁹/L.

In some embodiments, administration of the sustained-release dosage formof the invention to a human once daily for at least 16 weeks results ina mean decrease in mean baseline hemoglobin (Hgb) of no more than about15 g/L, no more than about 10 g/L, no more than about 8 g/L, or no morethan about 6 g/L.

In some embodiments, administration of the sustained-release dosage formof the invention to a human once daily for at least 16 weeks results ina mean decrease in mean baseline hemoglobin (Hgb) of about 0 to about 15g/L, about 5 to about 15 g/L, about 2 to about 12 g/L, or about 5 toabout 12 g/L.

In some embodiments, the platelet counts and hemoglobin levels aremeasured in patients having received no blood transfusions during thetreatment period.

Mean baseline platelet counts and mean baseline hemoglobin levels aretypically measured prior to the start of treatment.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, can also beprovided in combination in a single embodiment (while the embodimentsare intended to be combined as if written in multiply dependent form).Conversely, various features of the invention which are, for brevity,described in the context of a single embodiment, can also be providedseparately or in any suitable subcombination.

Methods

Another aspect of the present invention pertains to methods of treatinga JAK-associated disease or disorder in an individual (e.g., patient) byadministering to the individual in need of such treatment asustained-release dosage form of the invention. A JAK-associated diseasecan include any disease, disorder or condition that is directly orindirectly linked to expression or activity of the JAK, includingoverexpression and/or abnormal activity levels. A JAK-associated diseasecan also include any disease, disorder or condition that can beprevented, ameliorated, or cured by modulating JAK activity.

Examples of JAK-associated diseases include diseases involving theimmune system including, for example, organ transplant rejection (e.g.,allograft rejection and graft versus host disease).

Further examples of JAK-associated diseases include autoimmune diseasessuch as multiple sclerosis, rheumatoid arthritis, juvenile arthritis,psoriatic arthritis, type I diabetes, lupus, psoriasis, inflammatorybowel disease, ulcerative colitis, Crohn's disease, myasthenia gravis,immunoglobulin nephropathies, myocarditis, autoimmune thyroid disorders,chronic obstructive pulmonary disease (COPD), and the like. In someembodiments, the autoimmune disease is an autoimmune bullous skindisorder such as pemphigus vulgaris (PV) or bullous pemphigoid (BP).

Further examples of JAK-associated diseases include allergic conditionssuch as asthma, food allergies, eszematous dermatitis, contactdermatitis, atopic dermatitis (atropic eczema), and rhinitis. Furtherexamples of JAK-associated diseases include viral diseases such asEpstein Barr Virus (EBV), Hepatitis B, Hepatitis C, HIV, HTLV 1,Varicella-Zoster Virus (VZV) and Human Papilloma Virus (HPV).

Further examples of JAK-associated disease include diseases associatedwith cartilage turnover, for example, gouty arthritis, septic orinfectious arthritis, reactive arthritis, reflex sympathetic dystrophy,algodystrophy, Tietze syndrome, costal athropathy, osteoarthritisdeformans endemica, Mseleni disease, Handigodu disease, degenerationresulting from fibromyalgia, systemic lupus erythematosus, scleroderma,or ankylosing spondylitis.

Further examples of JAK-associated disease include congenital cartilagemalformations, including hereditary chrondrolysis, chrondrodysplasias,and pseudochrondrodysplasias (e.g., microtia, enotia, and metaphysealchrondrodysplasia).

Further examples of JAK-associated diseases or conditions include skindisorders such as psoriasis (for example, psoriasis vulgaris), atopicdermatitis, skin rash, skin irritation, skin sensitization (e.g.,contact dermatitis or allergic contact dermatitis). For example, certainsubstances including some pharmaceuticals when topically applied cancause skin sensitization. In some embodiments, co-administration orsequential administration of at least one JAK inhibitor of the inventiontogether with the agent causing unwanted sensitization can be helpful intreating such unwanted sensitization or dermatitis. In some embodiments,the skin disorder is treated by topical administration of at least oneJAK inhibitor of the invention.

In further embodiments, the JAK-associated disease is cancer includingthose characterized by solid tumors (e.g., prostate cancer, renalcancer, hepatic cancer, pancreatic cancer, gastric cancer, breastcancer, lung cancer, cancers of the head and neck, thyroid cancer,glioblastoma, Kaposi's sarcoma, Castleman's disease, uterineleiomyosarcoma, melanoma etc.), hematological cancers (e.g., lymphoma,leukemia such as acute lymphoblastic leukemia (ALL), acute myelogenousleukemia (AML) or multiple myeloma), and skin cancer such as cutaneousT-cell lymphoma (CTCL) and cutaneous B-cell lymphoma. Example CTCLsinclude Sezary syndrome and mycosis fungoides.

In some embodiments, the JAK inhibitors described herein, or incombination with other JAK inhibitors, such as those reported in U.S.Ser. No. 11/637,545, which is incorporated herein by reference in itsentirety, can be used to treat inflammation-associated cancers. In someembodiments, the cancer is associated with inflammatory bowel disease.In some embodiments, the inflammatory bowel disease is ulcerativecolitis. In some embodiments, the inflammatory bowel disease is Crohn'sdisease. In some embodiments, the inflammation-associated cancer iscolitis-associated cancer. In some embodiments, theinflammation-associated cancer is colon cancer or colorectal cancer. Insome embodiments, the cancer is gastric cancer, gastrointestinalcarcinoid tumor, gastrointestinal stromal tumor (GIST), adenocarcinoma,small intestine cancer, or rectal cancer.

JAK-associated diseases can further include those characterized byexpression of: JAK2 mutants such as those having at least one mutationin the pseudo-kinase domain (e.g., JAK2V617F); JAK2 mutants having atleast one mutation outside of the pseudo-kinase domain; JAK1 mutants;JAK3 mutants; erythropoietin receptor (EPOR) mutants; or deregulatedexpression of CRLF2.

JAK-associated diseases can further include myeloproliferative disorders(MPDs) such as polycythemia vera (PV), essential thrombocythemia (ET),primary myelofibrosis (PMF), chronic myelogenous leukemia (CML), chronicmyelomonocytic leukemia (CMML), hypereosinophilic syndrome (HES),systemic mast cell disease (SMCD), and the like. In some embodiments,the myeloproliferative disorder is myelofibrosis (e.g., primarymyelofibrosis (PMF) or post polycythemia vera/essential thrombocythemiamyelofibrosis (Post-PV/ET MF)). In some embodiments, themyeloproliferative disorder is post-essential thrombocythemiamyelofibrosis (Post-ET MF). In some embodiments, the myeloproliferativedisorder is post polycythemia vera myelofibrosis (Post-PV MF).

The present invention further provides methods of treating psoriasis orother skin disorders by administration of a topical formulationcontaining a compound of the invention.

In some embodiments, sustained-release formulation and dosage formsdescribed herein can be used to treat pulmonary arterial hypertension.

In some embodiments, the sustained-release formulation and dosage formsdescribed herein can be used to treat mast cell activation syndrome.

The present invention further provides a method of treatingdermatological side effects of other pharmaceuticals by administrationof the sustained-release dosage form of the invention. For example,numerous pharmaceutical agents result in unwanted allergic reactionswhich can manifest as acneiform rash or related dermatitis. Examplepharmaceutical agents that have such undesirable side effects includeanti-cancer drugs such as gefitinib, cetuximab, erlotinib, and the like.The dosage form of the invention can be administered in combination with(e.g., simultaneously or sequentially) the pharmaceutical agent havingthe undesirable dermatological side effect.

Further JAK-associated diseases include inflammation and inflammatorydiseases. Example inflammatory diseases include sarcoidosis,inflammatory diseases of the eye (e.g., iritis, uveitis, scleritis,conjunctivitis, or related disease), inflammatory diseases of therespiratory tract (e.g., the upper respiratory tract including the noseand sinuses such as rhinitis or sinusitis or the lower respiratory tractincluding bronchitis, chronic obstructive pulmonary disease, and thelike), inflammatory myopathy such as myocarditis, and other inflammatorydiseases. In some embodiments, the inflammation disease of the eye isblepharitis.

The sustained-release dosage forms herein can further be used to treatischemia reperfusion injuries or a disease or condition related to aninflammatory ischemic event such as stroke or cardiac arrest. Thesustained-release dosage forms described herein can further be used totreat endotoxin-driven disease state (e.g., complications after bypasssurgery or chronic endotoxin states contributing to chronic cardiacfailure). The sustained-release dosage forms described herein canfurther be used to treat anorexia, cachexia, or fatigue such as thatresulting from or associated with cancer. The sustained-release dosageforms described herein can further be used to treat restenosis,sclerodermitis, or fibrosis. The sustained-release dosage formsdescribed herein can further be used to treat conditions associated withhypoxia or astrogliosis such as, for example, diabetic retinopathy,cancer, or neurodegeneration. See, e.g., Dudley, A. C. et al. Biochem.J. 2005, 390(Pt 2):427-36 and Sriram, K. et al. J. Biol. Chem. 2004,279(19):19936-47. Epub 2004 Mar. 2, both of which are incorporatedherein by reference in their entirety. The sustained-release dosageforms described herein can be used to treat Alzheimer's disease.

The sustained-release dosage forms described herein can further be usedto treat other inflammatory diseases such as systemic inflammatoryresponse syndrome (SIRS) and septic shock.

The sustained-release dosage forms described herein can further be usedto treat gout and increased prostate size due to, e.g., benign prostatichypertrophy or benign prostatic hyperplasia.

Further JAK-associated diseases include bone resorption diseases such asosteoporosis, osteoarthritis. Bone resorption can also be associatedwith other conditions such as hormonal imbalance and/or hormonaltherapy, autoimmune disease (e.g. osseous sarcoidosis), or cancer (e.g.myeloma). The reduction of the bone resorption due to the JAK inhibitorscan be about 10%, about 20%, about 30%, about 40%, about 50%, about 60%,about 70%, about 80%, or about 90%.

In some embodiments, sustained-release dosage forms described herein canfurther be used to treat a dry eye disorder. As used herein, “dry eyedisorder” is intended to encompass the disease states summarized in arecent official report of the Dry Eye Workshop (DEWS), which defined dryeye as “a multifactorial disease of the tears and ocular surface thatresults in symptoms of discomfort, visual disturbance, and tear filminstability with potential damage to the ocular surface. It isaccompanied by increased osmolarity of the tear film and inflammation ofthe ocular surface.” Lemp, “The Definition and Classification of Dry EyeDisease: Report of the Definition and Classification Subcommittee of theInternational Dry Eye Workshop”, The Ocular Surface, 5(2), 75-92 April2007, which is incorporated herein by reference in its entirety. In someembodiments, the dry eye disorder is selected from aqueoustear-deficient dry eye (ADDE) or evaporative dry eye disorder, orappropriate combinations thereof. In some embodiments, the dry eyedisorder is Sjogren syndrome dry eye (SSDE). In some embodiments, thedry eye disorder is non-Sjogren syndrome dry eye (NSSDE).

In a further aspect, the present invention provides a method of treatingconjunctivitis, uveitis (including chronic uveitis), chorioditis,retinitis, cyclitis, sclieritis, episcleritis, or iritis; treatinginflammation or pain related to corneal transplant, LASIK (laserassisted in situ keratomileusis), photorefractive keratectomy, or LASEK(laser assisted sub-epithelial keratomileusis); inhibiting loss ofvisual acuity related to corneal transplant, LASIK, photorefractivekeratectomy, or LASEK; or inhibiting transplant rejection in a patientin need thereof, comprising administering to the patient atherapeutically effective amount of the compound of the invention, or apharmaceutically acceptable salt thereof.

Additionally, the sustained-release dosage forms of the invention,optionally in combination with other JAK inhibitors such as thosereported in U.S. Ser. No. 11/637,545, which is incorporated herein byreference in its entirety, can be used to treat respiratory dysfunctionor failure associated with viral infection, such as influenza and SARS.

As used herein, the term “individual,” “subject,” or “patient,” refersto a human, who can be fasted or un-fasted when the dosage form of theinvention is administered.

As used herein the term “pharmaceutically acceptable” refers to thosecompounds, materials, compositions and/or dosage forms, which are,within the scope of sound medical judgment, suitable for contact humanswithout excessive toxicity, irritation, allergic response and otherproblem complications commensurate with a reasonable benefit/risk ratio.

As used herein, the term “treating” or “treatment” refers to one or moreof (1) preventing the disease; for example, preventing a disease,condition or disorder in an individual who may be predisposed to thedisease, condition or disorder but does not yet experience or displaythe pathology or symptomatology of the disease; (2) inhibiting thedisease; for example, inhibiting a disease, condition or disorder in anindividual who is experiencing or displaying the pathology orsymptomatology of the disease, condition or disorder (i.e., arrestingfurther development of the pathology and/or symptomatology); and (3)ameliorating the disease; for example, ameliorating a disease, conditionor disorder in an individual who is experiencing or displaying thepathology or symptomatology of the disease, condition or disorder (i.e.,reversing the pathology and/or symptomatology) such as decreasing theseverity of disease.

Combination Therapies

One or more additional pharmaceutical agents such as, for example,chemotherapeutics, anti-inflammatory agents, steroids,immunosuppressants, as well as Bcr-Abl, Flt-3, RAF and FAK kinaseinhibitors such as, for example, those described in WO 2006/056399,which is incorporated herein by reference in its entirety, or otheragents can be used in combination with the sustained-release dosageforms described herein for treatment of JAK-associated diseases,disorders or conditions. The one or more additional pharmaceuticalagents can be administered to a patient simultaneously or sequentially.

Example chemotherapeutics include proteosome inhibitors (e.g.,bortezomib), thalidomide, revlimid, and DNA-damaging agents such asmelphalan, doxorubicin, cyclophosphamide, vincristine, etoposide,carmustine, and the like.

Example steroids include coriticosteroids such as dexamethasone orprednisone.

Example Bcr-Abl inhibitors include the compounds, and pharmaceuticallyacceptable salts thereof, of the genera and species disclosed in U.S.Pat. No. 5,521,184, WO 04/005281, and U.S. Ser. No. 60/578,491, all ofwhich are incorporated herein by reference in their entirety.

Example suitable Flt-3 inhibitors include compounds, and theirpharmaceutically acceptable salts, as disclosed in WO 03/037347, WO03/099771, and WO 04/046120, all of which are incorporated herein byreference in their entirety.

Example suitable RAF inhibitors include compounds, and theirpharmaceutically acceptable salts, as disclosed in WO 00/09495 and WO05/028444, both of which are incorporated herein by reference in theirentirety.

Example suitable FAK inhibitors include compounds, and theirpharmaceutically acceptable salts, as disclosed in WO 04/080980, WO04/056786, WO 03/024967, WO 01/064655, WO 00/053595, and WO 01/014402,all of which are incorporated herein by reference in their entirety.

In some embodiments, one or more of the compounds of the invention canbe used in combination with one or more other kinase inhibitorsincluding imatinib, particularly for treating patients resistant toimatinib or other kinase inhibitors.

In some embodiments, one or more JAK inhibitors of the invention can beused in combination with a chemotherapeutic in the treatment of cancer,such as multiple myeloma, and may improve the treatment response ascompared to the response to the chemotherapeutic agent alone, withoutexacerbation of its toxic effects. Examples of additional pharmaceuticalagents used in the treatment of multiple myeloma, for example, caninclude, without limitation, melphalan, melphalan plus prednisone [MP],doxorubicin, dexamethasone, and Velcade (bortezomib). Further additionalagents used in the treatment of multiple myeloma include Bcr-Abl, Flt-3,RAF and FAK kinase inhibitors. Additive or synergistic effects aredesirable outcomes of combining a JAK inhibitor of the present inventionwith an additional agent. Furthermore, resistance of multiple myelomacells to agents such as dexamethasone may be reversible upon treatmentwith a JAK inhibitor of the present invention. The agents can becombined with the present compounds in a single or continuous dosageform, or the agents can be administered simultaneously or sequentiallyas separate dosage forms.

In some embodiments, a corticosteroid such as dexamethasone isadministered to a patient in combination with at least one JAK inhibitorwhere the dexamethasone is administered intermittently as opposed tocontinuously.

In some further embodiments, combinations of sustained-release dosageforms with other therapeutic agents can be administered to a patientprior to, during, and/or after a bone marrow transplant or stem celltransplant.

In some embodiments, the additional therapeutic agent is fluocinoloneacetonide (Retisert®), or rimexolone (AL-2178, Vexol, Alcon).

In some embodiments, the additional therapeutic agent is cyclosporine(Restasis®).

In some embodiments, the additional therapeutic agent is acorticosteroid. In some embodiments, the corticosteroid istriamcinolone, dexamethasone, fluocinolone, cortisone, prednisolone, orflumetholone.

In some embodiments, the additional therapeutic agent is selected fromDehydrex™ (Holles Labs), Civamide (Opko), sodium hyaluronate (Vismed,Lantibio/TRB Chemedia), cyclosporine (ST-603, Sirion Therapeutics),ARG101(T) (testosterone, Argentis), AGR1012(P) (Argentis), ecabet sodium(Senju-Ista), gefarnate (Santen), 15-(s)-hydroxyeicosatetraenoic acid(15(S)-HETE), cevilemine, doxycycline (ALTY-0501, Alacrity),minocycline, iDestrin™ (NP50301, Nascent Pharmaceuticals), cyclosporineA (Nova22007, Novagali), oxytetracycline (Duramycin, MOLI1901,Lantibio), CF101(2S,3S,4R,5R)-3,4-dihydroxy-5-[6-[(3-iodophenyl)methylamino]purin-9-yl]-N-methyl-oxolane-2-carbamyl,Can-Fite Biopharma), voclosporin (LX212 or LX214, Lux Biosciences),ARG103 (Agentis), RX-10045 (synthetic resolvin analog, Resolvyx), DYN15(Dyanmis Therapeutics), rivoglitazone (DE011, Daiichi Sanko), TB4(RegeneRx), OPH-01 (Ophtalmis Monaco), PCS101 (Pericor Science), REV1-31(Evolutec), Lacritin (Senju), rebamipide (Otsuka-Novartis), OT-551(Othera), PAI-2 (University of Pennsylvania and Temple University),pilocarpine, tacrolimus, pimecrolimus (AMS981, Novartis), loteprednoletabonate, rituximab, diquafosol tetrasodium (INS365, Inspire), KLS-0611(Kissei Pharmaceuticals), dehydroepiandrosterone, anakinra, efalizumab,mycophenolate sodium, etanercept (Embrel®), hydroxychloroquine, NGX267(TorreyPines Therapeutics), actemra, gemcitabine, oxaliplatin,L-asparaginase, or thalidomide.

In some embodiments, the additional therapeutic agent is ananti-angiogenic agent, cholinergic agonist, TRP-1 receptor modulator, acalcium channel blocker, a mucin secretagogue, MUC1 stimulant, acalcineurin inhibitor, a corticosteroid, a P2Y2 receptor agonist, amuscarinic receptor agonist, an mTOR inhibitor, another JAK inhibitor,Bcr-Abl kinase inhibitor, Flt-3 kinase inhibitor, RAF kinase inhibitor,and FAK kinase inhibitor such as, for example, those described in WO2006/056399, which is incorporated herein by reference in its entirety.In some embodiments, the additional therapeutic agent is a tetracyclinederivative (e.g., minocycline or doxycline). In some embodiments, theadditional therapeutic agent binds to FKBP12.

In some embodiments, the additional therapeutic agent is an alkylatingagent or DNA cross-linking agent; an anti-metabolite/demethylating agent(e.g., 5-flurouracil, capecitabine or azacitidine); an anti-hormonetherapy (e.g., hormone receptor antagonists, SERMs, or aromotaseinhibitor); a mitotic inhibitor (e.g. vincristine or paclitaxel); antopoisomerase (I or II) inhibitor (e.g. mitoxantrone and irinotecan); anapoptotic inducers (e.g. ABT-737); a nucleic acid therapy (e.g.antisense or RNAi); nuclear receptor ligands (e.g., agonists and/orantagonists: all-trans retinoic acid or bexarotene); epigenetictargeting agents such as histone deacetylase inhibitors (e.g.vorinostat), hypomethylating agents (e.g. decitabine); regulators ofprotein stability such as Hsp90 inhibitors, ubiquitin and/or ubiquitinlike conjugating or deconjugating molecules; or an EGFR inhibitor(erlotinib).

In some embodiments, the additional therapeutic agent(s) are demulcenteye drops (also known as “artificial tears”), which include, but are notlimited to, compositions containing polyvinylalcohol, hydroxypropylmethylcellulose, glycerin, polyethylene glycol (e.g. PEG400), orcarboxymethyl cellulose. Artificial tears can help in the treatment ofdry eye by compensating for reduced moistening and lubricating capacityof the tear film. In some embodiments, the additional therapeutic agentis a mucolytic drug, such as N-acetyl-cysteine, which can interact withthe mucoproteins and, therefore, to decrease the viscosity of the tearfilm.

In some embodiments, the additional therapeutic agent includes anantibiotic, antiviral, antifungal, anesthetic, anti-inflammatory agentsincluding steroidal and non-steroidal anti-inflammatories, andanti-allergic agents. Examples of suitable medicaments includeaminoglycosides such as amikacin, gentamycin, tobramycin, streptomycin,netilmycin, and kanamycin; fluoroquinolones such as ciprofloxacin,norfloxacin, ofloxacin, trovafloxacin, lomefloxacin, levofloxacin, andenoxacin; naphthyridine; sulfonamides; polymyxin; chloramphenicol;neomycin; paramomycin; colistimethate; bacitracin; vancomycin;tetracyclines; rifampin and its derivatives (“rifampins”); cycloserine;beta-lactams; cephalosporins; amphotericins; fluconazole; flucytosine;natamycin; miconazole; ketoconazole; corticosteroids; diclofenac;flurbiprofen; ketorolac; suprofen; cromolyn; lodoxamide; levocabastin;naphazoline; antazoline; pheniramine; or azalide antibiotic.

The invention will be described in greater detail by way of specificexamples. The following examples are offered for illustrative purposes,and are not intended to limit the invention in any manner. Those ofskill in the art will readily recognize a variety of non-criticalparameters which can be changed or modified to yield essentially thesame results.

EXAMPLES Example 1 Sustained-Release and Immediate-Release Formulationsof Ruxolitinib Phosphate Formulation SR-2

A 25 mg sustained-release formulation of ruxolitinib phosphate wasprepared according to the following protocol. The formulation componentsare provided in Table 1a. Percentages are by weight.

TABLE 1a Component Function Percentage Ruxolitinib phosphate^(a) Activeingredient 12.2 Microcryrstalline cellulose, NF Filler 22.0Hypromellose, USP (Methocel K15M) Sustained release 4.0 matrix formerHypromellose, USP (Methocel K4M) Sustained release 16.0 matrix formerLactose monohydrate, NF Filler 42.3 Colloidal silicon dioxide, NFGlidant 1.0 Magnesium stearate, NF Lubricant 0.5 Stearic acid, NFLubricant 2.0 Total 100 ^(a)On a free base basis, conversion factor forphosphate salt to free base is 0.7575.

Protocol

Step 1. Add microcrystalline cellulose, ruxolitinib phosphate, lactosemonohydrate, and hypromelloses to a suitable blender and mix.Step 2. Transfer the mix from Step 1 to a suitable granulator and mix.Step 3. Add purified water while mixing.Step 4. Screen the wet granules from Step 3.Step 5. Transfer the granules from Step 4 into a suitable dryer and dryuntil LOD is no more than 3%.Step 6. Screen the granules from Step 5.Step 7. Mix colloidal silicon dioxide with granules in Step 6 in asuitable blender.Step 8. Mix stearic acid and magnesium stearate with the blend in Step 7and continue blending.Step 9. Compress the final blend in Step 8 on a suitable rotary tabletpress.

Formulation SR-1

An alternate 25 mg sustained-release formulation of ruxolitinibphosphate has been prepared as described below. The formulationcomponents are provided in Table 1b. Percentages are by weight.

TABLE 1b Component Function Percentage Ruxolitinib phosphate^(a) Activeingredient 12.2 Microcryrstalline cellulose, NF Filler 42.3Hypromellose, USP (Methocel K100LV) Sustained release 10.0 matrix formerHypromellose, USP (Methocel K4M) Sustained release 12.0 matrix formerLactose monohydrate, NF Filler 20.0 Colloidal silicon dioxide, NFGlidant 1.0 Magnesium stearate, NF Lubricant 0.5 Stearic acid, NFLubricant 2.0 Total 100 ^(a)On a free base basis, conversion factor forphosphate salt to free base is 0.7575.

Protocol

Step 1. Add microcrystalline cellulose, ruxolitinib phosphate, lactosemonohydrate, and hypromelloses to a suitable blender and mix.Step 2. Transfer the mix from Step 1 to a suitable granulator and mix.Step 3. Add purified water while mixing.Step 4. Screen the wet granules from Step 3.Step 5. Transfer the granules from Step 4 into a suitable dryer and dryuntil LOD is no more than 3%.Step 6. Screen the granules from Step 5.Step 7. Mix colloidal silicon dioxide with granules in Step 6 in asuitable blender.Step 8. Mix stearic acid and magnesium stearate with the blend in Step 7and continue blending.Step 9. Compress the final blend in Step 8 on a suitable rotary tabletpress.

Formulation C—Immediate Release

Immediate-release dosage forms of ruxolitinib phosphate can be obtainedcommercially in 5, 10, 15, 20, and 25 mg doses as the drug productJakafi® (ruxolitinib phosphate (tablets)) (NDA no. N202192). Thecommercially available dosage forms are the same as used in the Phase 3COMFORT-1 and COMFORT-II studies.

Example 2 Bioavailability Study of Sustained Release Formulation

A relative bioavailability study of the sustained release and immediateformulations of ruxolitinib phosphate was conducted in healthy adultvolunteers. Subjects in the fasted state were given a single oral doseof immediate-release formulation (25 mg, see Example 1) or a single oraldose of sustained-release formulation (25 mg, see Example 1). Plasmaconcentrations of ruxolitinib were measured and are compared in FIG. 1.Table 2a provides comparative pharmacokinetic (PK) data.

TABLE 2a* C_(max) T_(max) C_(12 h) t_(1/2) AUC_(0-t) AUC_(0-∞) Cl/FTreatment n (nM) (h) (nM) C_(max)/C_(12 h) (h) (nM * h) (nM * h) (L/h)25 mg IR 9 1100 ± 332  0.94 ± 0.46 45.6 ± 38.1 40 ± 24 2.8 ± 0.72 4340 ±1990 4350 ± 1990 22.8 ± 10.3 1060 0.86 32.1 33 2.7 3930 3940 20.7 25 mgSR-1 8  333 ± 76.1  2.4 ± 0.98  121 ± 46.8 3.0 ± 1.0 5.3 ± 1.8  3110 ±840  3180 ± 864  27.2 ± 6.72 325 2.2 114 2.9 5.1 3020 3090 26.4 25 mgSR-2 8 394 ± 126 2.9 ± 1.6  104 ± 43.2 4.7 ± 3.1 6.1 ± 2.1  3520 ± 12603740 ± 1400 24.6 ± 9.02 377 2.5 96.5 3.9 5.8 3330 3520 23.2 P-Valuesfrom a Crossover ANOVA of Log-Transformed Data <0.0001 0.0003 — —<0.0001 0.040 0.070   0.070 Geometric Mean Relative Bioavailability and90% Cl (Reference = IR) SR-1 vs. IR 30.4% 74.7% 76.2% 25.4-36.4%62.2-89.7% 63.1-92.0% SR-2 vs. IR 35.2% 82.5% 86.7% 29.5-42.2%68.7-99.1% 71.8-105% *PK values are provided as mean ± SD and geometricmean

Description of Bioavailability Study

This study was performed to evaluate pharmacokinetic performance of tworuxolitinib phosphate sustained release (SR) formulations compared tothe ruxolitinib phosphate immediate release (IR) tablets. The study wasconducted as a 3-period study in which each subject received the IRtablets, the SR-1 tablets and the SR-2 tablets, all in fasted state. Alltreatments were administered as a single dose in one tablet. Ninehealthy subjects enrolled in this study received IR tablets in Period 1,and 8 subjects continued on the study were randomized into 2 sequencesto receive SR-1 and SR-2 tablets in Period 2 and Period 3.

The 9 subjects enrolled in the study received a single dose of IRtablets, SR-1 tablets and SR-2 tablets, according to the randomizationschedule (see Table 2b). Dosing was administered orally after at least10 hour overnight fast, and a standardized meal was served approximately3 hours after administration. A washout period of 7 days (not less than5 days) was instituted between the treatment periods.

Blood samples for determination of plasma concentrations of ruxolitinibwere collected at 0, 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 16, 24 and 36hours post-dose using lavender top (K2EDTA) Vacutainer® tubes. Nosamples were collected for determination of urine concentrations ofruxolitinib.

TABLE 2b Randomization Schedule for Study Sequence Period 1 Period 2Period 3 Subject 1 25 mg IR 25 mg SR-1 25 mg SR-2 tablet 102*, 103, 106,tablet tablet fasted fasted 107, 109 fasted 2 25 mg IR 25 mg SR-2 25 mgSR-1 tablet 101, 104, 105, tablet tablet fasted fasted 108 fasted*Subject withdrew from the study before the start of Period 2

Plasma and urine samples were shipped to Incyte Corporation fordetermination of ruxolitinib concentrations. The plasma samples wereassayed by validated, GLP, LC/MS/MS methods with a linear range of 1 to1000 nM.

All PK blood samples were collected within 5 minutes of their scheduledtime, and therefore the schedule times relative to the time of doseadministration were used for all pharmacokinetic analyses.

Standard non-compartmental pharmacokinetic methods were used to analyzethe ruxolitinib plasma concentration data using Phoenix WinNonlinversion 6.0 (Pharsight Corporation, Mountain View, Calif.). Thus,C_(max) and T_(max) were taken directly from the observed plasmaconcentration data. The absorption lag time (T_(lag)) was defined as thesampling time immediately preceding that corresponding to the firstmeasurable (non-zero) concentration. The terminal-phase disposition rateconstant (λ_(z)) was estimated using a log-linear regression of theconcentration data in the terminal disposition phase, and t_(1/2) wasestimated as ln(2)/λ_(z). AUC_(0-t) was estimated using the lineartrapezoidal rule for increasing concentrations and the log-trapezoidalrule for decreasing concentrations, and the total AUC_(0-∞) wascalculated as AUC_(0-t)+C_(t)/λ_(z). The oral-dose clearance (Cl/F) wasestimated as Dose/AUC_(0-∞), and the terminal-phase volume ofdistribution (V_(z)/F) was estimated as Dose/[AUC_(0-∞)*λ_(z)].

The log-transformed pharmacokinetic parameters were compared among thetreatments using a 2-factor ANOVA with the fixed factor for treatmentand random factor for subject. The relative bioavailability of thefasted administration of the SR formulations (test treatments) comparedto fasted administration of the IR tablets (reference treatment) wereestimated using the geometric mean relative bioavailability and 90%confidence intervals for C_(max), AUC_(0-t) and AUC_(0-∞), which werecalculated from the adjusted means (least square means) from the ANOVA.All statistical analyses were performed using SAS version 9.1 (SASInstitute, Inc., Cary, N.C.).

Example 3 Sustained-Release Clinical Trial

A Phase 2 clinical trial was carried out in patients with myelofibrosis(MF). A total of 41 subjects were enrolled, and spleen volume and totalsymptom scores were obtained at baseline. Tablets of sustained-releaseformulation SR-2 (see Example 1) were administered to fasted patients.All patients were treated with 25 mg once daily doses for 8 weeks. After8 weeks, depending upon the clinical response, the investigator wasallowed to (a) maintain the same dose of SR-2, (b) increase the dose to50 mg once daily, (c) increase the dose to alternating doses of 25 mgand 50 mg, dosed once daily, or (d) switch to treatment with theimmediate release formulation.

Data related to spleen volume and total symptom scores are provided inFIGS. 2 and 3 together with comparative data from the COMFORT-1 Studywhere patients were dosed only with the immediate-release formulation.See Example A below for details of the COMFORT-1 Study. As can be seenin FIGS. 2 and 3, treatment with the 25 mg sustained-release formulationwas nearly as effective as the immediate-release formulation in theCOMFORT-I study. Percentages of spleen volume responders in COMFORT-1 asshown in FIG. 2 were 39.4% and 43.9% at 12 weeks and 24 weeks,respectively. The percentage of spleen volume responders in thesustained-release study at week 16 was 28.9%. Similarly, the percentagesof total symptom score responders in COMFORT-1 as shown in FIG. 3 were46.3% and 45.9% at 12 weeks and 24 weeks, respectively. The percentageof total symptom score responders in the sustained release study asshown in FIG. 3 was 36.8%.

Data relating to mean platelet count and mean hemoglobin levels (Hgb) ispresented below in Table 3 together with comparative data from theCOMFORT-I study. As can be seen from the data, the mean change frombaseline platelet count in the SR patients was about half of what wasobserved in COMFORT-I. Similar results are seen for hemoglobin levels.Data is shown as mean±SD.

TABLE 3 Sustained- COMFORT-I COMPFORT-I Release Study Study ParameterStudy (active) (placebo) N 41 155 154 Mean Baseline 274 ± 193  321 ± 202280 ± 152 platelet count, ×10⁹/L Mean Baseline Hgb in 105 ± 17  108 ± 20106 ± 22  patient with no transfusions, g/L Week 16 Week 12 Week 12 Meantotal daily dose, 34   30.4  0 mg Mean change from −65 ± 101 −131 ± 143−9 ± 75 baseline platelet count, ×10⁹/L Mean change from −7.4 ± 13.3−13.2 ± 15.5  0.3 ± 11.5 baseline Hgb, g/L

Example 4 Comparison of Steady State Plasma Concentrations ofRuxolitinib between SR and IR Formulations in MF Patients

Steady state plasma concentrations of ruxolitinib in myelofibrosis (MF)patients receiving repeating 25 mg doses are compared in FIG. 4 betweensustained release (SR) and immediate release (IR) formulations.Comparative pharmacokinetic parameters are provided below in Table 4a.

TABLE 4a* Formula and C_(max) T_(max) C_(min) t_(1/2) AUC_(0-τ)AUC_(0-t) Cl/F Regimen n (nM) (h) (nM) (h) (nM * h) (nM * h) (L/h) SR 39397 ± 175 2.35 ± 1.75   32 ± 40.6 7.33 ± 3.73 3650 ± 2450 2060 ± 114032.0 ± 18.0 25 mg QD^(a)  368 1.88 — 6.42 3020 1810 27.1 IR 25 mg 271481 ± 575  0.83 ± 0.45 47 ± 54 1.94 ± 0.50 4363 ± 2066 4148 ± 1885 22.7± 10.1 BID^(b) 1374 0.74 — 1.88 3949 3778 20.7 (1A) IR 25 mg QD^(b) 61417 ± 150  0.84 ± 0.38 0 ± 0 1.60 ± 0.36 3567 ± 777  3291 ± 604  23.9 ±5.5  (2A) 1410 0.78 — 1.57 3494 3243 23.4 IR 25 mg 7 1650 ± 506  0.79 ±0.49  85 ± 102 1.96 ± 0.59 4939 ± 2566 4444 ± 1918 19.9 ± 8.1  BID^(b)1578 0.68 43 1.90 4463 4120 18.3 (2C) SR 25 mg 8 394 ± 126 2.9 ± 1.6 —6.1 ± 2.1 3740 ± 1400 — 24.6 ± 9.02 single dose^(c)  377 2.5  5.8  352023.2 ^(a)From SR study described in Example 3 ^(b)From IR studydescribed below (Description of Open-Label Study in MF patients)^(c)From single dose study described in Example 2 (AUC_(0-τ) will beAUC_(0-∞)) *Values are mean ± SD and geometric mean.Description of Open-Label Study in ME patients

General Description

This was an open-label study exploring the safety, tolerability, andefficacy of ruxolitinib, administered orally to patients with primarymyelofibrosis (PMF) and post polycythemia vera/essential thrombocythemiamyelofibrosis (Post-PV/ET MF). The study was comprised of 3 parts: Part1-dose escalation and expansion, bid dosing, Part 2—alternative dosingschedules (A, B and C), and Part 3-three independent patient groups(Group I, II and III). Eight dose regimens were evaluated in 3 parts.The Part 1 evaluated two dose levels of 25 mg bid and 50 mg bid, Part 2studied five dose regimens of 10 mg bid, 25 mg bid, 25 mg qd, 50 mg qdand 100 mg qd and Part 3 assessed six dose regimens of 10 mg bid, 15 mgbid, 25 mg bid, 50 mg qd, 100 mg qd and 200 mg qd. A total of 154subjects were enrolled; 32 subjects enrolled in Part 1, 29 subjects inPart 2 and 93 subjects in Part 3. See Table 4b (qd=once per day;bid=twice per day).

In Part 1, the pharmacokinetic blood samples were collected at pre-doseand 0.5, 1, 1.5, 2, 4, 6 and 9 hours post-dose on Days 1 and 15 of Cycle1 and at pre-dose on Day 1 of Cycles 2 and 3, using lavender top(K3EDTA) Vacutainer® tubes. In Part 2, the pharmacokinetic samples werecollected at pre-dose and 0.5, 1, 1.5, 2, 4, 6 and 9 hours post-dose onDay 15 of Cycle 1 and at pre-dose on Day 1 of Cycles 2 and 3. In Part 3,the pharmacokinetic samples were collected at pre-dose and 2 hours afteradministration of the morning dose of ruxolitinib on Day 15 of Cycle 1and Day 1 of Cycles 2 and 3.

Plasma concentration data from Cycle 1 for subjects in Part 1 and 2 wereused for non-compartmental analysis while all plasma concentration datawere used for population PK analysis.

Following fasting, oral, first-dose or multiple-dose administration ofruxolitinib phosphate tablets, the drug was absorbed rapidly, typicallyattaining peak plasma concentrations within 0.3 to 2 hours afteradministration in all subjects. Plasma concentrations subsequentlydeclined in a monophasic or biphasic fashion.

The mean C_(max) and AUC increased approximately linearly proportionalto dose from 10 mg to 100 mg. The pharmacokinetics of ruxolitinib in MFpatients was similar to that in healthy volunteers.

Detailed Description of Clinical Trial

This multicenter, open-label, non-randomized, dose escalation clinicalstudy was conducted by M.D. Anderson Cancer Center, Houston, Tex. andMayo Clinic, Rochester, Minn., according to Protocol INCB 18424-251, and154 patients with PMF or Post-PV/ET MF were enrolled and received atleast a single dose according to the study plan in Table 4b. The studywas comprised of 3 parts: Part 1—dose escalation and expansion cohort,bid dosing, Part 2—alternative dosing schedules (A, B and C), and Part3-three independent patient groups (Group I, II and III). Schedules A, Band C in Part 2 were once daily (qd) dosing regimens, low dose regimenof 10 mg bid and induction/maintenance regimen, respectively. Part 3 wasstudied in three separate groups of patients to further evaluate thesafety and efficacy of selected starting dose levels and to explore dosemodification on an individual patient basis as appropriate. Dose holdand withdrawal for safety were defined in terms of platelet count andabsolute neutrophil count (ANC) while provision for dose increase wasprovided based on inadequate efficacy defined by change in spleen size.

Ruxolitinib phosphate tablets (5 and 25 mg) were administered as oraldoses with water in an outpatient setting. Doses ranged from 10 mg bidto 50 mg bid, and from 25 mg qd to 200 mg qd. The individual patientparticipation was expected to be approximately 12-24 months; patientsmight continue on therapy indefinitely if they did not meet any of thewithdrawal criteria, did not have disease progression and are receivingsome clinical benefit.

In Part 1, the pharmacokinetic blood samples were collected at pre-doseand 0.5, 1, 1.5, 2, 4, 6 and 9 hours post-dose on Days 1 and 15 of Cycle1 and at pre-dose on Day 1 of Cycles 2 and 3, using lavender top(K3EDTA) Vacutainer® tubes. In Part 2, the pharmacokinetic samples werecollected at pre-dose and 0.5, 1, 1.5, 2, 4, 6 and 9 hours post-dose onDay 15 of Cycle 1 and at pre-dose on Day 1 of Cycles 2 and 3. In Part 3,the pharmacokinetic samples were collected at pre-dose and 2 hours afteradministration of the morning dose on Day 15 of Cycle 1 and Day 1 ofCycles 2 and 3.

TABLE 4b Part Schedule Dose Regimen 1 A 25 mg bid 1 B 50 mg bid 2 A 25mg qd 2 A 50 mg qd 2 A 100 mg qd 2 B 10 mg bid 2 C 25 mg bid 3 I 50 mgqd 3 I 10 mg qd 3 I 25 mg bid 3 II 100 mg qd 3 II 200 mg qd 3 III 10 mgbid 3 III 15 mg bid

Plasma samples were shipped to Incyte Corporation and assayed by avalidated, GLP, LC/MS/MS method with a linear range of 1 to 1000 nM anda limit of quantification of 1 nM.

Generally, the actual time post-dose was used for pharmacokineticanalyses. However, the dose information on Cycle 1 Day 15 for patientsin Part 2 and three additional patients in Part 1 were not collected.The nominal time were used for pharmacokinetic analyses for thesepatients. The dose information on Cycle 1 Day 15 for four additionalpatients was questionable. Hence, the nominal time were used for thesepatients also. Plasma concentrations at 12 hours post dose for bid or 24hours post dose for qd on Cycle 1, Day 15 were imputed by sample atpre-dose on Cycle 1 Day 15 to calculate steady state AUC_(0-τ).

Standard noncompartmental pharmacokinetic methods were used to analyzethe ruxolitinib plasma concentration data using WinNonlin version 6.0(Pharsight Corporation, Mountain View, Calif.). Thus, C_(max) andT_(max) were taken directly from the observed plasma concentration data.For single dose, the terminal-phase disposition rate constant (λ_(z))was estimated using a log-linear regression of the concentration data inthe terminal disposition phase, and t_(1/2) was estimated asln(2)/λ_(z). AUC_(0-t) was estimated using the linear-trapezoidal rulefor increasing concentrations and the log-trapezoidal rule fordecreasing concentrations, and the total AUC_(0-∞) was calculated asAUC_(0-t)+C_(t)/λ_(z). The oral-dose clearance (Cl/F) was estimated asDose/AUC_(0-∞), and the terminal-phase volume of distribution (V_(z)/F)was estimated as Dose/[AUC_(0-∞)*λ_(z)].

For the multiple-dose data, λ_(z) was estimated using a log-linearregression of the concentration data in the terminal disposition phase,and t_(1/2) was estimated as ln(2)/λ_(z). The AUC over one dosinginterval (AUC_(0-12h) for q12 h administration, or AUC_(0-24h) for q24 hadministration) was estimated using the linear trapezoidal rule forincreasing concentrations and the log-trapezoidal rule for decreasingconcentrations. The Cl/F was estimated as Dose/AUC, and V_(z)/F wasestimated as Dose/[AUC*λ_(z)]. Additionally, the C_(min) and AUC_(0-t)(Area under the steady-state plasma concentration-time curve from timezero to the time of the last sample obtained) were calculated for themultiple-dose data.

The PK parameters of ruxolitinib were summarized for each dose groupusing descriptive statistics, and the log-transformed ruxolitinib PKparameters were compared among the dose groups using a 1-factor analysisof variance. The dose-proportionality of C_(max) and AUC was evaluatedusing a power function regression model (eg, C_(max)=α·Dose β).

The pharmacokinetics of ruxolitinib in MF patients was similar to thatin healthy volunteers.

Example 5 Comparative Efficacy of Sustained Release and ImmediateRelease Formulations

Enlarged spleen is a common and prominent symptom of myelofibrosis.Reduction in spleen volume serves as a measure for assessing theeffectiveness of a given treatment. Table 5a reports the mean reductionin spleen volume in MF patients enrolled in the sustained-release study(See Example 3) at 16 weeks of treatment, while Table 5b reports themean reduction in spleen volume in MF patients enrolled in the COMFORT-Istudy (immediate release, see Comparative Example A) at 24 weeks oftreatment. As can be seen from the data, both the sustained-release andimmediate release treatment regimens were effective in reducing spleenvolume.

TABLE 5a Spleen Volume (cm³) from Sustained-Release Study Percent Changefrom Baseline to Week 16 (%) n 40 Mean −22.3 STD 20.79 Median −21.7(MIN, MAX) (−64.6, 43.6)

TABLE 5b Spleen Volume (cm³) from COMFORT I (Immediate-Release)Treatment Group Percent Change from Baseline To Week 12 (%) To Week 24(%) Ruxolitinib Placebo Ruxolitinib Placebo (N = 155) (N = 154) (N =155) (N = 154) n 148 132 139 106 Mean −32.0 8.4 −31.6 8.1 STD 15.5814.61 18.92 15.31 Min −74.4 −26.2 −75.9 −46.4 Median −31.7 6.1 −33.0 8.5Max 3.8 64.6 25.1 48.8

Effectiveness of a treatment regimen in an MF patient can also beassessed by Total Symptom Score. In calculating Total Symptom Score,symptoms of MF were assessed using a symptom diary (modified MFSAF v2.0diary) where subjects recorded answers to queries regarding MF symptomson a handheld device. Symptoms assessed included filling upquickly/early satiety, abdominal discomfort, abdominal pain, inactivity,night sweats, itching, and bone/muscle pain.

Table 5c reports the Total Symptom Score results in thesustained-release study (see Example 3) while Table 5d reports the TotalSymptom Score results in the COMFORT-I study (immediate-release, seeComparative Example A) at 24 weeks. As can be seen from the data, boththe SR and IR regimens were effective at treating MF in patients.

TABLE 5c Total Symptom Scores from Sustained-Release Study PercentChange from Baseline to Week 16 n 38 Mean −50.4 STD 31.16 Median −48.6(MIN, MAX) (−100.0, 12.7)

TABLE 5D Total Symptom Score from COMFORT I Study (Immediate-Release)Treatment Group Ruxolitinib Placebo Ruxolitinib Placebo Percent Changefrom Baseline To Week 16 To Week 24 (N = 155) (N = 154) (N = 155) (N =154) n 140 124 129 103 Mean −40.5 37.8 −46.1 41.8 STD 54.31 93.92 48.5599.26 Min −100.0 −82.5 −100.0 −100.0 Median −51.1 12.7 −56.2 14.6 Max292.5 464.8 108.3 511.6

Example 6 Comparison of Adverse Events in Patients Enrolled in theSustained-Release Study and COMFORT-I (Immediate-Release) Study

Data for adverse events relating to anemia, thrombocytopenia,neutropenia, and all Grade 3 or higher adverse events are compared forthe sustained-release and COMFORT-I (immediate-release) studies (seeExample 3 and Comparative Example A for descriptions of the studies) inTable 6a. Adverse events are graded according to CTCAE criteria whichcan be found online at ctep.cancer.gov/protocolDevelopment/electronicapplications/ctc.htm orevs.nci.nih.gov/ftpl/CTCAE/CTCAE_(—)4.03_(—)2010-06-14_QuickReference_(—)5x7.pdf.A Grade 3 adverse event generally corresponds to a reaction that issevere or medically significant, but not immediately life-threateningwhere hospitalization or prolongation of hospitalization is indicatedand where the reaction is disabling to the extent of limiting self care.Higher Grades are 4 (life-threatening requiring urgent intervention) and5 (death). For anemia, Grade 3 corresponds to Hgb<8.0 g/dL; <4.9 mmol/L;<80 g/L, where a transfusion is indicated. For thrombocytopenia(decreased platelet count), Grade 3 corresponds to <50,000-25,000/mm³;<50.0-25.0×10⁹/L. The sustained-release data was evaluated for patientsover the course of 16 weeks. Mean duration of exposure to ruxolitinib inthe COMFORT-I was approximately 242 days. Typically, the majority ofhematologic adverse events occur within the first few months of therapyas observed in the COMFORT-I study.

As can be seen from the data in Table 6a, adverse events relating toanemia, thrombocytopenia, neutropenia, and all events that wereconsidered Grade 3 or higher occurred less frequently in thesustained-release study compared with the COMFORT-I immediate-releasestudy.

TABLE 6a Percent of Patients with Selected Grade 3 or Higher AdverseEvents COMFORT I COMFORT I Adverse Event SR Study Ruxolitinib PlaceboAll ≧Grade 3 Adverse 17.1%   47.1% 44.4% Events Anemia 0% 15.5% 4.6%Thrombocytopenia 2.4%   8.4% 2.0% Neutropenia 0% 1.3% 0.7%

The occurrence of blood-related adverse events are further compared inTables 6b and 6c which report the number and percentage of patients inthe studies exhibiting certain reactions including anemia andthrombocytopenia. The sustained-release data was evaluated for patientsover the course of 16 weeks. Mean duration of exposure to ruxolitinib inthe COMFORT-I study was approximately 242 days. The vast majority ofhematologic adverse events occur within the first few months of therapyas observed in the COMFORT-I study. As can be seen from the data in thetables, the number and percentage of patients exhibiting blood-relatedadverse events is lower in the sustained-release study. Additionally,the severity of the adverse events is lesser in the sustained-releasestudy.

TABLE 6b Treatment-Related Haematologic Adverse Events By Organ Class,Preferred Term, and Maximum Severity in the Sustained - Release Study(First 16 Weeks)* Ruxolitinib MedDRA System Organ Class/ (N = 41) MedDRAPreferred Term Mi Mo Se LT Any Number (%) of Subjects With Any 16 9 7 032 (78.0) Adverse Events Blood and lymphatic system disorders 3 4 1 0 8(19.5) Anaemia 0 2 0 0 2 (4.9) Thrombocytopenia 3 2 1 0  6 (14.6) *Mi(mild), Mo (moderate), Se (severe), LT (life threatening)

TABLE 6c Treatment-Related Adverse Events By MedDRA System Organ Class,Preferred Term, and Maximum Severity in COMFORT I* MedDRA SystemRuxolitinib Placebo Organ Class/ (N = 155) (N = 151) MedDRA PreferredTerm Mi Mo Se LT FT Any Mi Mo Se LT FT Any Number (%) of Subjects 20 5855 9 9 151 (97.4)  23 58 53 4 10 148 (98.0)  With Any Adverse EventsBlood and lymphatic 20 28 27 10 0 85 (54.8) 18 21 15 2 0 56 (37.1)system disorders Anaemia 4 20 16 8 0 48 (31.0) 4 10 7 0 0 21 (13.9)Thrombocytopenia 21 19 11 2 0 53 (34.2) 6 5 2 1 0 14 (9.3)  *Mi (mild),Mo (moderate), Se (severe), LT (life threatening), FT (fatal)

Comparative Example A COMFORT I Clinical Trial—Immediate ReleaseFormulation

A Phase 3 clinical trial was completed showing efficacy of ruxolitinibin myelofibrosis patients. In this double-blind trial, patients withintermediate-2 or high risk myelofibrosis were randomly assigned totwice-daily oral, immediate-release (see Example 1) ruxolitinib (155patients) or placebo (154 patients). The starting dose of ruxolitinibdepended on the baseline platelet count: 15 mg twice daily for aplatelet count of 100×109 to 200×109 per liter and 20 mg twice daily fora count that exceeded 200×109 per liter. The dose was adjusted for lackof efficacy or excess toxicity. The primary end point was the proportionof patients with a reduction in spleen volume of 35% or more at 24weeks, assessed by means of magnetic resonance imaging. Secondary endpoints included the durability of response, changes in symptom burden(assessed by the total symptom score), and overall survival.

The proportion of patients with a reduction of 35% or more in spleenvolume at week 24 (primary end point) was 41.9% in the ruxolitinib groupas compared with 0.7% in the placebo group. The proportion of patientswith a reduction of 50% or more in the total symptom score from baselineto week 24, a pre-specified secondary end point, was significantlyhigher in the ruxolitinib group than in the placebo group (45.9% vs.5.3%; odds ratio, 15.3; 95% CI, 6.9 to 33.7; P<0.001).

The study showed that ruxolitinib was associated with reductions insplenomegaly and symptoms that are prominent manifestations ofmyelofibrosis and appeared to be associated with an improvement inoverall survival. Additionally, the most common toxic effects of anemiaand thrombocytopenia were generally managed with dose modification.Details regarding this study are provided in Verstovsek, S., et al. “Adouble-blind, placebo-controlled trial of ruxolitinib formyelofibrosis,” N. Eng. J. Med., 2012, Mar. 1:366(9):799-807, which isincorporated herein by reference in its entirety.

Comparative Example B COMFORT II Clinical Trial—Immediate ReleaseFormulation

A Phase 3 clinical trial was completed showing the superiority ofruxolitinib treatment in myelofibrosis patients compared with bestavailable therapy. Continuous ruxolitinib therapy, as compared with thebest available therapy, was associated with marked and durablereductions in splenomegaly and disease-related symptoms, improvements inrole functioning and quality of life, and modest toxic effects.

Myelofibrosis patients were randomly assigned, in a 2:1 ratio, toreceive ruxolitinib or the best available therapy, which included anycommercially available agents (as monotherapy or in combination) or notherapy at all and which could be changed during the treatment phase.The starting dose of ruxolitinib tablets was 15 mg twice daily of animmediate release formulation (See Example 1) if the baseline plateletcount was 200×109 per liter or less and 20 mg orally twice daily if thebaseline platelet count was greater than 200×109 per liter.

The primary end point was a reduction of 35% or more in spleen volumefrom baseline at week 48. At week 48, most of the patients in theruxolitinib group had a reduction in spleen volume. Only patients in theruxolitinib group met the criterion for the primary end point, at leasta 35% reduction in spleen volume from baseline at 48 weeks (28%, vs. 0%in the group receiving the best available therapy; P<0.001). Patients inthe ruxolitinib group, as compared with patients receiving the bestavailable therapy, had improved quality of life and role functioning. Atweek 48, patients receiving ruxolitinib had marked reductions inmyelofibrosis associated symptoms, including appetite loss, dyspnea,fatigue, insomnia, and pain, whereas patients receiving the bestavailable therapy had worsening symptoms.

Thrombocytopenia and anemia occurred more frequently in the patientsreceiving ruxolitinib than in those receiving the best availabletherapy, but these events were generally manageable with dosemodifications, transfusions of packed red cells, or both. Additionaldetails of the study are provided in Harrison, C. et al., “JAKinhibition with ruxolitinib versus best available therapy formyelofibrosis,” N. Eng. J. Med., 2012, Mar. 1; 366(9):787-98 which isincorporated herein by reference in its entirety.

Various modifications of the invention, in addition to those describedherein, will be apparent to those skilled in the art from the foregoingdescription. Such modifications are also intended to fall within thescope of the appended claims. Each reference, including all patent,patent applications, and publications, cited in the present applicationis incorporated herein by reference in its entirety.

What is claimed is:
 1. A sustained-release dosage form comprising atleast one active ingredient which is ruxolitinib, or a pharmaceuticallyacceptable salt thereof, wherein said ruxolitinib, or pharmaceuticallyacceptable salt thereof, is present in said dosage form in an amount ofabout 10 to about 60 mg on a free base basis.
 2. The sustained-releasedosage form of claim 1 wherein said ruxolitinib, or pharmaceuticallyacceptable salt thereof, is present in said dosage form in an amount ofabout 25 mg on a free base basis.
 3. The sustained-release dosage formof claim 1, wherein said active ingredient is ruxolitinib phosphate. 4.The sustained-release dosage form of claim 1, wherein administration ofsaid dosage form to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 700 nM or less.
 5. Thesustained-release dosage form of claim 1, wherein administration of saiddosage form to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 200 to about 700 nM.
 6. Thesustained-release dosage form of claim 1, wherein administration of saiddosage form to a human results in a mean peak plasma concentration(C_(max)) of ruxolitinib of about 300 to about 400 nM.
 7. Thesustained-release dosage form of claim 1, wherein administration of saiddosage form to a human state results in a mean time to peak plasmaconcentration (T_(max)) of ruxolitinib of about 1.5 hours or more. 8.The sustained-release dosage form of claim 1, wherein administration ofsaid dosage form to a human results in a mean time to peak plasmaconcentration (T_(max)) of ruxolitinib of about 1.5 hours to about 5hours.
 9. The sustained-release dosage form of claim 1, whereinadministration of said dosage form to a human results in a ratio of meanpeak plasma concentration (C_(max)) to mean 12-hour plasma concentration(C_(12h)) of ruxolitinib of about 10 or less.
 10. The sustained-releasedosage form of claim 1, wherein administration of said dosage form to ahuman results in a ratio of mean peak plasma concentration (C_(max)) tomean 12-hour plasma concentration (C_(12h)) of ruxolitinib of about 4 orless.
 11. The sustained-release dosage form of claim 1, whereinadministration of said dosage form to a human results in a meanhalf-life (t_(1/2)) of from about 3.5 hours to about 11 hours.
 12. Thesustained-release dosage form of claim 1, wherein administration of saiddosage form to a human results in a mean half-life (t_(1/2)) of fromabout 4 hours to about 8 hours.
 13. The sustained-release dosage form ofclaim 1, wherein administration of a single dose of said dosage form toa human results in mean bioavailability (AUC_(0-∞)) of ruxolitinib ofabout 3000 to about 4000 nM*h.
 14. The sustained-release dosage form ofclaim 1, wherein administration of a single dose of said dosage form toa human results in mean bioavailability (AUC_(0-∞)) of ruxolitinib ofabout 3100 to about 3800 nM*h.
 15. The sustained-release dosage form ofclaim 1 comprising one or more cellulosic ethers.
 16. Thesustained-release dosage form of claim 15 comprising hydroxypropylmethylcellulose.
 17. The sustained-release dosage form of claim 15comprising from about 10% to about 30% by weight of hydroxypropylmethylcellulose.
 18. The sustained-release dosage form of claim 1 whichis in the form of a tablet or capsule.
 19. The sustained-release dosageform of claim 1, comprising 25 mg of ruxolitinib on a free base basis,or a pharmaceutically acceptable salt thereof, wherein administration toa patient results in a mean ruxolitinib plasma level of about 75 toabout 500 nM for at least about 8 hours.
 20. The sustained-releasedosage form of claim 19 comprising 12-13% ruxolitinib phosphate byweight on a free base basis and 19-13% by weight of one or morehypromelloses.
 21. The sustained-release dosage form of claim 19comprising 12.2% ruxolitinib phosphate by weight on a free base basisand 20% by weight or 22% by weight of one or more hypromelloses.
 22. Thesustained-release dosage form of claim 1, wherein administration to apatient results in a ruxolitinib plasma level of about 75 to about 500nM for at least about 8 hours.
 23. The sustained-release dosage form ofclaim 1, wherein administration to a patient results in a ruxolitinibplasma level of about 75 to about 500 nM for at least about 12 hours.24. The sustained-release dosage form of claim 1 comprising 25 mgruxolitinib phosphate on a free base basis, wherein administration ofsaid dosage form to a patient for at least 16 weeks results in a meandecrease in mean base platelet count of no more than about 100×10⁹/L.25. The sustained-release dosage form of claim 24 wherein administrationof said dosage form to a patient for at least 16 weeks results in a meandecrease in mean base platelet count of no more than about 80×10⁹/L. 26.The sustained-release dosage form of claim 24 wherein administration ofsaid dosage form to a patient for at least 16 weeks results in a meandecrease in mean base platelet count of no more than about 60×10⁹/L. 27.The sustained-release dosage form of claim 24 wherein administration ofsaid dosage form to a patient for at least 16 weeks results in a meandecrease in mean base platelet count of no more than about 40×10⁹/L. 28.The sustained-release dosage form of claim 1, comprising 25 mgruxolitinib phosphate on a free base basis, wherein administration ofsaid dosage form to a patient for at least 16 weeks results in a meandecrease in mean hemoglobin of no more than about 15 g/L.
 29. Thesustained-release dosage form of claim 28 wherein administration of saiddosage form to a patient for at least 16 weeks results in a meandecrease in mean hemoglobin of no more than about 10 g/L.
 30. Thesustained-release dosage form of claim 28 wherein administration of saiddosage form to a patient for at least 16 weeks results in a meandecrease in mean hemoglobin of no more than about 8 g/L.
 31. Thesustained-release dosage form of claim 28 wherein administration of saiddosage form to a patient for at least 16 weeks results in a meandecrease in mean hemoglobin of no more than about 6 g/L.
 32. Thesustained-release dosage form of claim 1 comprising (1) ruxolitinibphosphate, or a pharmaceutically acceptable salt thereof, (2)microcrystalline cellulose, (3) hypromellose, (4) lactose monohydrate,(5) colloidal silicon dioxide, (6) magnesium stearate, and (7) stearicacid.
 33. A method of treating a disease associated with JAK activity ina patient in need thereof, comprising administering the dosage form ofclaim 1 to said patient, wherein said disease is selected from anautoimmune disease, a skin disorder, allograft rejection, graft versushost disease, multiple sclerosis, rheumatoid arthritis, juvenilearthritis, type I diabetes, lupus, inflammatory bowel disease, Crohn'sdisease, myasthenia gravis, immunoglobulin nephropathies, myocarditis,autoimmune thyroid disorder, a viral disease, Epstein Barr Virus (EBV),Hepatitis B, Hepatitis C, HIV, HTLV 1, Varicella-Zoster Virus (VZV),Human Papilloma Virus (HPV), cancer, a myeloproliferative disorder, aninflammatory disease, an inflammatory disease of the eye, iritis,uveitis, scleritis, conjunctivitis, an inflammatory disease of therespiratory tract, an inflammatory disease of the upper respiratorytract, an inflammatory disease of the lower respiratory tract, aninflammatory myopathy, myocarditis, ischemia reperfusion or a disorderrelated to an ischemic event, anorexia or cachexia resulting from orassociated with cancer, fatigue resulting from or associated withcancer, a bone resorption disease, or mast cell activation syndrome. 34.The method of claim 33, wherein said autoimmune disease is bullous skindisorder.
 35. The method of claim 34, wherein said bullous skin disorderis pemphigus vulgaris (PV) or bullous pemphigoid (BP).
 36. The method ofclaim 33, wherein said skin disorder is atopic dermatitis, psoriasis,skin sensitization, skin irritation, skin rash, contact dermatitis orallergic contact sensitization.
 37. The method of claim 36, wherein saidskin disorder is psoriasis.
 38. The method of claim 33, wherein saidmyeloproliferative disorder (MPD) is polycythemia vera (PV), essentialthrombocythemia (ET), primary myelofibrosis (PMF), post-polycythemiavera myelofibrosis (post PV-MF), post-essential thrombocythemiamyelofibrosis (post ET-MF), chronic myelogenous leukemia (CML), chronicmyelomonocytic leukemia (CMML), hypereosinophilic syndrome (HES), orsystemic mast cell disease (SMCD).
 39. The method of claim 38, whereinsaid disease is primary myelofibrosis (PMF), post-polycythemia veramyelofibrosis (post PV-MF), post-essential thrombocythemia myelofibrosis(post ET-MF), polycythemia vera (PV), or essential thrombocythemia (ET).40. The method of claim 38, wherein said disease is primarymyelofibrosis (PMF), post-polycythemia vera myelofibrosis (post PV-MF),or post-essential thrombocythemia myelofibrosis (post ET-MF).
 41. Themethod of claim 33, wherein said cancer is a solid tumor, myeloma,prostate cancer, renal cancer, hepatic cancer, breast cancer, lungcancer, thyroid cancer, Kaposi's sarcoma, Castleman's disease,pancreatic cancer, hematological cancer, lymphoma, leukemia, multiplemyeloma, skin cancer, cutaneous T-cell lymphoma or cutaneous B-celllymphoma.
 42. The method of claim 33, wherein said bone resorptiondisease is osteoporosis, osteoarthritis, bone resorption associated withhormonal imbalance, bone resorption associated with hormonal therapy,bone resorption associated with autoimmune disease, or bone resorptionassociated with cancer.
 43. The method of claim 33, wherein said oraldosage form is administered once daily.